scholarly journals IDENTIFICATION OF METHICILLIN RESISTANT STAPHYLOCOCCUS AUREUS USING TOUCHDOWN PCR AND PHENOTYPIC METHODS FROM PATIENTS AND HOSPITALS ENVIRONMENTS IN DIFFERENT IRAQI CITIES

2021 ◽  
Vol 52 (6) ◽  
pp. 1356-1364
Author(s):  
A. M. Abd Zaid ◽  
N. J. Kandala

The study was aimed to evaluate the prevalence of MRSA in some Iraqi hospitals and determine the most powerful methods for identification of MRSA, in order to achieve the, 278 samples were collected from different hospitals in Iraq in various intervals, 204 out of 287 were identified as Staphylococcus aureus by conventional cultural methods and microscopic characteristics and 177 isolates are identified as MRSA by using HiCrome MeReSa Agar Base medium, but 154 of 177 (87%) isolates are methicillin resistance in sensitivity test. MRSA isolates were highly resistant to β-lactam antibiotics and considered multidrug resistant (MDR) in percent of (94.9%). Touchdown PCR used to identify the isolates, 97.05% were identified as Staphylococcus aureus, while 80.88%  as MRSA.                  

2021 ◽  
pp. 50-66
Author(s):  
Iyekhoetin Matthew Omoruyi ◽  
Helen Inikpi Ajayi

Aim: The aim of this study was to determine the prevalence of Shigella spp and methicillin resistant Staphylococcus aureus in some selected commercially processed ready-to-eat snail (Archachatina marginata) and edible worm (Rhynchophorus phoenicis). Study Design: Cross-sectional analysis. Place and Duration of Study: Samples were obtained from vendors along the Benin-Sapele express road, South-South Nigeria, over a two months period (November to December, 2019). Methodology: The enumeration of total heterotrophic counts, total Shigella counts and total Staphylococcus aureus counts were done using nutrient agar, Salmonella Shigella agar and mannitol salt agar respectively. All isolates were further identified by their cultural, morphological and biochemical characteristics. Methicillin resistant Staphylococcus aureus were identified using methicillin-oxacillin agar, while the antibiogramic profile of selected isolates and their multidrug resistant profile were done according to the CSLI guideline. Results: The mean heterotrophic bacterial counts (THCs) obtained in Rhynchophorus phoenicis ranged from 0.00 × 10³ ± 0.00 cfu/g to 500.00 × 10³ ± 0.00 cfu/g, while the mean THCs obtained in Archachatina marginata ranged from 13.3 × 10³ ± 1.15 cfu/g to 500 × 10³ ± 0.00 cfu/g. The total Staphylococcus aureus obtained in Rhynchophorus phoenicis was between 0.00 × 10³ ± 0.00 cfu/g to 294 × 10³ ± 4.7 cfu/g and the Shigella bacteria counts from 0.00 × 10³ ± 0.00 cfu/g to 258 × 10³ ± 14.64 cfu/g. The antibiogram of Staphylococcus aureus isolated from both Rhynchophorus phoenicis and Archachatina marginata, found the majority of isolates (96.67%) to be resistant to ceftazidime and cefuroxime, while 83.33% of the isolates were sensitive to ofloxacin. All Staphylococcus isolates were resistant to ceftriaxone, cloxacillin and amoxycillin clavulanate. The multidrug resistant index recorded was between 37.5 and 100. All Staphylococcus (9) isolates tested for methicillin resistance was observed to be positive. Conclusion: This study demonstrated that RTE vended meat products sold in along Benin City By-pass, contains Shigella and methicillin resistant Staphylococcus species, which are potential foodborne pathogens and efforts should be made at eliminating them from these vended meat products.


Author(s):  
Seighazi Regina Egege ◽  
Nedie Patience Akani ◽  
Chidiebele Emmanuel Ikechukwu Nwankwo

Aims/Objective: Methicillin-Resistant Staphylococcus aureus (MRSA) is a dominant cause of severe healthcare-associated (HA) infections but has recently emerged as Community-Associated (CA-MRSA) and Livestock-Associated MRSA (LA-MRSA). This study was carried out to detect the presence of Methicillin-Resistant S. aureus in Ready-to-Eat (RTE) Corbiculid heterodont in Bayelsa state, Nigeria. Place of Study: This study was conducted in Yenagoa, Southern Ijaw and Sagbama Local Government Areas, Bayelsa State, Nigeria. Methodology: Fifty-three samples (dried, fried and stewed) of the RTE C. heterodont were collected and subjected to standard microbiological procedures. Results: Seventy Staphylococcus spp. were isolated, consisting of 65 (92.86%) S. aureus and 5 (7.14%) coagulase negative Staphylococcus (CONS). Out of the 65 S. aureus isolates, 53.8% were MRSA. Susceptibility pattern of MRSA showed a decreasing trend of resistance in the order: Amoxacillin, Ampicillin-Cloxacillin, Cefoxitin, and Oxacillin (100%) > Streptomycin (82.9%) >Ceftriaxone (40%) >Cefuroxime (31.4%), > Ciprofloxacin and Trimethoprim-Sulfamethoxazole (17.1%) > Gentamycin and Pefloxacin (8.6%) > Erythromycin (2.9%).Out of the 65 S. aureus isolates, 62 (95.4%) had Multidrug Resistance Index > 0.2 while 3 (4.6%) had 0.2 as their Multidrug Resistance Index. The MRSA isolates were 100% Multidrug Resistant (MDR) while the Methicillin-Sensitive S. aureus (MSSA) were 90% MDR. Molecular characterization of ten S. aureus isolates confirmed 80% S. aureus isolates and 20% CONS (S. sciuri and S. warneri).  Polymerase chain reaction (PCR) and Agarose gel-electrophoresis were used to detect the presence of mecA gene, a gene that confers methicillin resistance. The mecA gene with size 500bp was identified in 50% of MRSA. This study reported presence of MRSA and MDR S. aureus in C. heterodont sold in Bayelsa state, Nigeria. This could be the avenue for the spread of MRSA. Public health awareness campaigns should be advocated to enhance hygienic practices in the handling and distribution of food.


Author(s):  
Maria Rukan ◽  
Humaira Jamil ◽  
Habib Ali Bokhari ◽  
Aamer Ali Khattak ◽  
Allah Nawaz Khan ◽  
...  

Abstract Objective: To isolate and characterise multidrug resistant strains of Staphylococcus aureus from healthcare workers who are at potential risk of nosocomial infections. Methods: The observational, cross-sectional study was conducted from November 2014 to April 2015 at different hospitals of Haripur and Abbottabad, Pakistan, and comprised ward and operation theatre staff. The isolates were identified on the basis of microbiological and biochemical tests and further confirmed by polymerase chain reaction. Disc diffusion method was used for antibiotic sensitivity testing, and panton valentine leukocidin and methicillin resistance mecA genes were detected using polymerase chain reaction. Results: Of 208 isolates, 108(52%) were from the ward staff and 100(48%) were from the operation theatre staff. Overall, 167(80.3%) isolates were positive for Staphylococcus aureus, and 75(36%) were methicillin-resistant Staphylococcus aureus. The number of antibiotic-resistant isolates was 75(45%) cefoxitin, 60(36%) ofloxacin, 152(91%) erythromycin, 52(31%) doxycycline, 127(76%) lincomycin, 53(32%) amoxicillin-clavulanate, 67(40%) ciprofloxacin, and 89(53%) ceftriaxone. Conclusion: A high number of hospital staff, including those working in operation theatres, were found to be carrying methicillin-resistant Staphylococcus aureus and multidrug resistant strains in their nasal passage that may be a source of infection to patients. Key Words: Methicillin resistance, Nosocomial infections, Vancomycin, MecA gene, Pvl gene. Continuous....


Author(s):  
Rahima Touaitia ◽  
Soumia Bektache ◽  
Nafissa Boutefnouchet ◽  
Abdelghani Djahoudi ◽  
Mohamed Bachtarzi

ABSTRACTObjective: The purpose of this study is to investigate the methicillin resistance gene, and some virulence factors in methicillin-resistant Staphylococcusaureus (MRSA) isolates by polymerase chain reaction (PCR).Methods: This study has included 12 MRSA isolates. All isolates were previously identified as S. aureus by a standard microbiological procedure, anda detection of methicillin resistance was realized by phenotypic methods. Following genomic DNA extraction, the presence of gyrA, mecA, lukPV, andtst genes was analyzed by duplex PCR. All retained S. aureus species have been found to contain gyrA gene.Results: Ten stains have been found to harbor mecA gene indicating it’s responsibility for methicillin resistance in those strains. Among the 12 strains,six of which were found to be Panton-Valentine leukocidine positive while none of which has tst gene encoding the toxic shock syndrome toxin.Conclusion: The pathogenesis of MRSA infections is related to the expression of a wide variety of virulence factors.Keywords: Methicillin-resistant Staphylococcus aureus, Panton-Valentine leukocidine, Toxic shock syndrome toxin 1, Multidrug resistance, mecA.


Author(s):  
VinodKumar C. S. ◽  
Sonika Prasad ◽  
Satish Patil ◽  
Aditya Reddy V. ◽  
Ankita Jha ◽  
...  

Background: The role of public transport as reservoirs of antibiotic-resistant staphylococci was determined.Methods: 200 swabs were collected from 50 public buses (urban and rural) circulating in Davangere, Karnataka. Swabs collected were inoculated on Blood agar, Mannitol salt agar and MacConkey agar plates. After incubation for 24-48 hours, plates were examined for the growth of Staphylococcus aureus. Anti-microbial susceptibility test was performed using oxacillin 1ug disc to detect methicillin resistance as per CSLI guidelines.Results: Out of 40 Staphylococcus aureus isolated 35 isolates were resistant to more than two classes of antibiotics, hence multidrug resistant Staphylococcus aureus. Out of 35 MDR isolates, 18 were resistant to oxacillin and cefoxitin. Minimum inhibitory concentration test revealed that out of 35 MDR isolates, 18 isolates had MIC value of ≥ 4µg/ml.Conclusions: The recovery methicillin-resistant Staphylococcus aureus from public transport system implies a potential risk for transmission of these bacteria in community.


Pathogens ◽  
2021 ◽  
Vol 10 (4) ◽  
pp. 427
Author(s):  
Martyna Kasela ◽  
Agnieszka Grzegorczyk ◽  
Bożena Nowakowicz-Dębek ◽  
Anna Malm

Nursing homes (NH) contribute to the regional spread of methicillin-resistant Staphylococcus aureus (MRSA). Moreover, residents are vulnerable to the colonization and subsequent infection of MRSA etiology. We aimed at investigating the molecular and phenotypic characteristics of 21 MRSA collected from the residents and personnel in an NH (Lublin, Poland) during 2018. All MRSA were screened for 20 genes encoding virulence determinants (sea-see, eta, etb, tst, lukS-F-PV, eno, cna, ebpS, fib, bbp, fnbA, fnbB, icaADBC) and for resistance to 18 antimicrobials. To establish the relatedness and clonal complexes of MRSA in NH we applied multiple-locus variable-number tandem-repeat fingerprinting (MLVF), pulse field gel electrophoresis (PFGE), multilocus sequence typing (MLST) and staphylococcal cassette chromosome mec (SCCmec) typing. We identified four sequence types (ST) among two clonal complexes (CC): ST (CC22) known as EMRSA-15 as well as three novel STs—ST6295 (CC8), ST6293 (CC8) and ST6294. All tested MRSA were negative for sec, eta, etb, lukS-F-PV, bbp and ebpS genes. The most prevalent gene encoding toxin was sed (52.4%; n = 11/21), and adhesins were eno and fnbA (100%). Only 9.5% (n = 2/21) of MRSA were classified as multidrug-resistant. The emergence of novel MRSA with a unique virulence and the presence of epidemic clone EMRSA-15 creates challenges for controlling the spread of MRSA in NH.


Pathogens ◽  
2021 ◽  
Vol 10 (4) ◽  
pp. 430
Author(s):  
Wichai Santimaleeworagun ◽  
Praewdow Preechachuawong ◽  
Wandee Samret ◽  
Tossawan Jitwasinkul

Methicillin-resistant Staphylococcus aureus (MRSA) is mostly found in Thailand in the hospital as a nosocomial pathogen. This study aimed to report the genetic characterization of a clinical community-acquired MRSA (CA-MRSA) isolate collected from hospitalized patients in Thailand. Among 26 MRSA isolates, S. aureus no. S17 preliminarily displayed the presence of a staphylococcal cassette chromosome mec (SCCmec) type IV pattern. The bacterial genomic DNA was subjected to whole-genome sequencing. Panton–Valentine leukocidin (PVL) production, virulence toxins, and antibiotic resistance genes were identified, and multi-locus sequence typing (MLST) and spa typing were performed. The strain was matched by sequence to MLST type 2885 and spa type t13880. This strain carried type IV SCCmec with no PVL production. Five acquired antimicrobial resistance genes, namely blaZ, mecA, Inu(A), tet(K), and dfrG conferring resistance to β-lactams, lincosamides, tetracycline, and trimethoprim, were identified. The detected toxins were exfoliative toxin A, gamma-hemolysin, leukocidin D, and leukocidin E. Moreover, there were differences in seven regions in CR-MRSA no. S17 compared to CA-MRSA type 300. In summary, we have reported the ST2885-SCCmec IV CA-MRSA clinical strain in Thailand for the first time, highlighting the problem of methicillin resistance in community settings and the consideration in choosing appropriate antibiotic therapy.


2014 ◽  
Vol 33 (10) ◽  
pp. e252-e259 ◽  
Author(s):  
Cilmara P. Garcia ◽  
Juliana F. Rosa ◽  
Maria A. Cursino ◽  
Renata D. Lobo ◽  
Carla H. Mollaco ◽  
...  

2005 ◽  
Vol 21 (3) ◽  
pp. 123-128 ◽  
Author(s):  
John G Gums ◽  
Benjamin J Epstein

Background: Staphylococcus aureus is a frequent cause of infections involving the bloodstream, skin and soft tissue, and lungs in hospitalized patients. These isolates are often multidrug resistant and represent a major therapeutic challenge. Objective: To explore the susceptibilities of S. aureus to nafcillin/oxacillin, a surrogate for methicillin resistance, and the fluoroquinolones and determine whether a relationship might exist between fluoroquinolone use and the prevalence of methicillin-resistant S. aureus (MRSA). Methods: To date, 353 institutions throughout the US and Puerto Rico have enrolled in the Antimicrobial Resistance Management (ARM) Program, and data have been submitted on nearly 5 million isolates of S. aureus. Isolates submitted from 1990 through 2002 were reviewed for sensitivity to nafcillin/oxacillin, clindamycin, erythromycin, and the fluoroquinolones ciprofloxacin and levofloxacin. Results: From 1990 through 2002 inclusive, susceptibility to nafcillin/oxacillin nationally was 64.9% (n = 360,460), ranging from 62.2% in the North Central and Northeast US to 72.8% in the Southwest. Nationally, S. aureus isolates were more resistant to levofloxacin (41.4%, n = 123,868) than ciprofloxacin (38.7%, n = 256,178). The greatest change in susceptibility of S. aureus to nafcillin/oxacillin and ciprofloxacin occurred concurrently from 1998 to 2002, which may implicate fluoroquinolone use with increasing rates of MRSA infection. Conclusions: Resistance to methicillin and the fluoroquinolones has increased in concert during the past 5 years. Collectively, data from the ARM Program, along with several other investigations, support a role of fluoroquinolone use in the emergence of MRSA. These observations, along with increasing resistance among gram-positive and gram-negative pathogens, underscore the need for judicious use of fluoroquinolones.


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